Plants are cellular tissue laden organisms having a genetic code for each plant within each species and plant variety. Within these genetic codes are variations or alterations which affect growth rates, yield and disease resistance.
Plants provide a large source of food, clothing, building supplies, paper products and medicines not to mention landscaping and beauty.
Rainforests contain no less than 60% of all higher plant species known on earth and they provide all that is needed for human survival, including remedies for disease. Their highly complex molecular structures often surpass the imagination of the chemical scientist and cannot easily be reproduced in the laboratory. More than 25% of all prescription drugs in Organisation for Economic Cooperation and Development (OECD) countries (contrasted with 60% in Eastern Europe) prove to consist of unmodified or slightly altered higher plant products. Natural drugs and medicinal plants, along with other non-timber forest products, already yield an important economic value. These few examples should make one realize how much modern drug delivery depends on sustainability and how vulnerable it is to the exhaustion of natural resources.
Plants are the source of many of our most important pharmaceuticals. Despite this, we know little about optimizing the production of these valuable secondary products in whole plants or cell and tissue cultures. Cultural practices to optimize pharmaceutical production in field or greenhouse grown plants have not been rigorously determined or have been of little benefit in increasing levels of the desired compounds. Considerable effort has been made to generate plant-derived pharmaceuticals economically in plant cell or tissue culture, with relatively few successes. As a result there is an apparent need to naturally stimulate growth and reproduction of these valuable plant species. The secret or key to continuing growth of such genetically complex plants will most likely occur by stimulation of growth factors within the plant's own tissues.
Commercially plants and plant products generate many hundreds of billions of dollars of commercial activity per year.
World demand for plant products is increasing very rapidly. The world demand for paper in 1997 was expected to increase by 50% by the year 2010. This places a huge demand on the timber industry which is concurrently seeing a surge in world demand for lumber products.
Trees like all of our plant products are renewable and in order to keep pace with increasing demands, faster growing and maturing trees are needed to avoid rapid deforestation on a worldwide basis.
Plants generally are taken somewhat for granted particularly in their role of influencing climate changes. Singularly no other species has a more positive role in affecting the global environment.
US patent publication 2005/0125161 A1 entitled “Differentially-Expressed Conifer cDNAS, and Their Use In Improving Somatic Embryogenesis” assigned to Institute of Paper Science and Technology provides a useful insight into current trends in coniferous trees and discloses a relational database of cDNA molecules, including those corresponding to Loblolly Pine Major Intrinsic Protein (MIP), which are differentially expressed during plant embryogenesis. The invention further related to the use of DNA arrays for evaluating gene expression in somatic and zygotic embryos. The invention encompassed related nucleic acids, proteins, antigens, and antibodies derived from these cDNAs as well as the use of such molecules for the staging, characterization, and manipulation of plant embryogenesis, in particular conifer embryogenesis. The cDNAs and related nucleic acids, proteins, antigens, and antibodies derived from these cDNAs are useful in the design, selection, and cultivation of improved crops, specifically including coniferous trees, which provide raw materials for paper and wood products.
Similarly, in US 2003/0074697 A1 entitled “Cotton Plants with Improved Cotton Fiber Characteristics and Methods for Producing Cotton Fibers From These Cotton Plants”, the inventors extensively studied the mechanisms of fiber elongation and formation in cotton plants from the viewpoints of molecular biology to improve the characteristics of cotton fibers. As a result, they found that this purpose can be attained by introducing a gene coding for endoxyloglucan transferase, which is deeply associated with the cell elongation and greatly expressed in the cotton fibers and ovule tissues at the cotton fiber elongation stage, or a gene coding for catalase or peroxidase, which is a hydrogen peroxide eliminating enzyme, into cotton plants and over-expressing these genes in the cotton fiber cells.
The result is a finer cotton fiber with a resultant higher yield. In this patent these benefits are achieved in an early stage by detection of a positive hybridization signal only from cDNA probe prepared from the ovules on the fifth day of flowering.
In US 2005/0044592 entitled “Plant Growth Modulation” teaches the use of one or more genes, encoding a protein of the elongator complex to modulate plant growth wherein there results an over expression of the DRL-1 gene to stimulate growth of leaves and roots, the subject matter of this publication being incorporated herein by reference in its entirety.
As in the other patents, stimulation occurs at the embryonic or early stage of plant development while the resultant growth modulation can occur throughout the life of the plant.
To better understand the fundamental aspects of the present invention the complexities of plants generally should be appreciated. In the background of US 2005/0044592 a summary of plant development is recited which reports findings of a variety of plant scientists which is repeated below.
Plants develop mainly post-germination from an embryo with a rudimentary body plan. The embryonic apical-basal axis is delineated by apical meristems that determine the future growth direction of the organism. The embryonic radial axis determines the identity and arrangement of tissues in concentric layers. During development pattern formation, growth and differentiation are overlapping rather than consecutive events. These processes are reiterated throughout the life cycle upon formation of every new organ. Axis formation is the basis for pattern formation within the whole plant body, an organ or even a tissue.
In Arabidopsis, leaves initiate post-germination at specific positions at the periphery of the shoot apical meristem according to a radial pattern imposed by the plant hormone auxin (Reinhardt et al., 2000). The repression of the homeobox gene SHOOT MERISTEMLESS and the activation of the myb gene ASYMMETRIC (AS) are crucial for leaf initiation (Long et al., 1996; Byrne et al., 2000). AS imposes a dorsi-ventral asymmetry upon the radial symmetry of the leaf primordium (Byrne et al., 2000). Dorsal identity in the leaf blade is promoted by the PHABULOSA and PHAVOLUTA transcription factors (TF) (McConnell et al., 2001) and ventral identity by the YABBY and KANADI TFs (Siegfried et al., 1999; Sawa et al., 1999; Kerstetter et al., 2001). Four tissues are specified along the dorsi-ventral axis: the upper epidermis and palissade parenchyma with dorsal identity, the spongy parenchyma and the lower epidermis with ventral identity.
In the primary root the radial axis of the radicle (embryonic root) is reinforced by positional information that originates from the top to the bottom, i.e. from mature cells to initial cells (van den Berg et al., 1995) and polar auxin transport (Sabatini et al., 1999). Tissues are arranged in concentric layers: the epidermis, the cortex, the endodermis, the pericycle and the vascular bundle. SCARECROW and SHORT ROOT are important genes for cortex specification (Scheres et al., 1995; Di Laurenzio et al., 1996), TORNADO 1 & 2 are important for epidermis specification (Cnops et al., 2000). Pattern formation in the primary root epidermal cell layer results in root hair cell files alternating with non-hair cell files which are formed at the anticlinal wall of two underlying cortex cells (Dolan et al., 1993, 1994). The gaseous hormone ethylene and auxin positively regulate root hair cell identity (Masucci et al., 1996). TRANSPARENT TESTA GLABRA1 and CAPRICE are positive regulators of root hair cell identity; GLABRA2 is a negative regulator (Di-Cristina et al., 1996; Wada et al., 1997; Walker et al., 1999).
The shoot apical meristem is essential for the formation of the vegetative plant body. Regulated cell division activity and changes in the orientation of cell plates precede the initiation of leaf primordia. Growth of leaf primordia occurs mainly along the length (proximo-distal axis) and width (centro-lateral axis) direction and is restricted along the thickness (dorsi-ventral axis) direction because of pattern formation in tissue layers. Early growth processes in leaves occur mainly by anticlinal cell divisions leading to the sheet-like structure of the blade. These growth processes are coupled with dorsi-ventral pattern formation (Siegried et al., 1999; McConnell et al., 2001; Eshed et al., 2001). Late growth occurs by cell expansion processes (Tsuge et al., 1996; Kim et al., 1998). Pattern formation in lateral growth results in the distinction between lamina and petiole (van der Graaff et al., 2000). Restriction of growth determines the final shape and size of the leaf organ. At flower induction, the SAM changes identity to an inflorescence meristem of which the structure and activity resembles that of the SAM except it produces floral meristems as lateral organs instead of leaf primordia. The onset of cell division in plants and animals is controlled at the G1/S transition of the cell cycle by the retinoblastoma protein that in a hypo-phosphorylated state binds and inactivates the general transcription factors E2F. Upon a mitogenic signal, such sucrose or cytokinin activated cyclin D/CDK complexes hyper-phosphorylate retinoblastoma and derepress E2F. By preventing cell cycle entry into S-phase, retinoblastoma plays a role in cell differentiation as well (de Jager and Murray, 1999). The cross-talk between cell cycle progression and developmental programs is a new and exciting area of research and the first reports have been published (Gaudin et al., 2000; De Veylder et al., 2001). Regulation of gene expression at the transcriptional level is an important and universal mechanism of controlling developmental programs. Classes of specific TFs recognize upstream promoter boxes in specific sets of genes. Through direct or indirect interaction with the general TFs the RNA polymerase II (RNAPII) transcription initiation complex is either activated or repressed. The specific TFs are activated by environmental or developmental stimuli that are transduced from the cell plasma membrane into the nucleus. Evidence in yeast and humans is accumulating that the control of expression of sets of genes is also mediated by the process of transcription elongation. The RNAPII transcription elongation complex forms the unfolded structure of transcribing nucleosomes (Walia et al., 1998). The elongation reaction is stimulated by a large variety of factors of which some prevent pausing or stalling of the RNAPII complex and others model the chromatin for transcription. The degree of chromatin condensation is modulated by histone acetyltransferases and deacetylases (Walia et al., 1998; Wittschieben et al., 1999). Elongating RNAPII holoenzyme co-purified with a multisubunit complex, Elongator, whose stable interaction is dependent on the hyperphosphorylated state of the RNAPII carboxy-terminal domain (Otero et al., 1999). The elongator complex consists of two subcomplexes: one consists of ELP1 (Otero et al., 1999), ELP2, a WD40 repeat protein (Fellows et al., 2000) and ELP3, a histone acetyltransferase (Wittschieben et al., 1999), the other one of ELP4, ELP5, and ELP6 (Krogan and Greenblatt, 2001; Winkler et al., 2001). Most components of Elongator are well conserved from yeast to man (Hawkes et al., 2001). Phenotypes of elpA mutants in yeast were slow growth adaptation, slow gene activation and temperature sensitivity and demonstrated that the ELP genes play a role in the activation of inducible genes in the adaptation to new growth conditions (Wittschieben et al., 1999; Otero et al., 1999; Fellows et al., 2000; Krogan and Greenblatt, 2001; Winkler et al., 2001). Mutations in man in one of the Elongator components cause familial dysautonomia, a well-known disorder (Hawkes et al., 2001). We identified the DEFORMED ROOT AND LEAF1 (DRL1) gene, a homolog of the yeast TOT4/KT112 gene (Butler et al., 1994; Frohloff et al., 2001). TOT genes were identified in search of mutants resistant to the Kluyveromyces lactis toxin zymocin. TOT1, TOT2, and TOT3 are isoallelic to ELP1, ELP2 and ELP3 and hence TOT equals elongator. TOT4/KT112 encodes a protein that associated with the elongator complex (Frohloff et al., 2001). The tot4 mutant displays similar phenotypes as deficient elongator mutants, in addition to slow growth, G1 cell cycle delay and hypersensitivity to Calcofluor White and caffeine. The inventors in US 2005/0044592 demonstrated that in higher plants DRL1 is important for pattern formation and growth processes.
The above related findings demonstrate that plants undergo a systemic response via a form of cross talk or cellular communication. This finding is consistent with a similar cellular communication found in mammals. In each organism be it a plant or mammal, cellular stimulation can result in a release of proteins and other chemical compositions relating to growth factors.
In attempts to activate such growth stimulations U.S. Pat. No. 5,819,467 entitled “Method of Stimulating Plant Growth” a conductive helical coil was spaced around a stem of a growing plant to stimulate growth by inducing an electromotive force or EMF.
Similarly in Canadian patent application CA 2 375 695 entitled “An Invention to Enhance Plant Growth and Germination” proposed growth and germination of some species of plants may be enhanced by exposure to a static magnetic field wherein permanent magnets were placed in a bank or array near the plants.
The present invention also has the object of stimulating plant growth and accelerating seed germination which is summarized as follows.